Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720, USA.
Nanoscale. 2011 Aug;3(8):3027-39. doi: 10.1039/c1nr10280j. Epub 2011 Jul 6.
This review is a practical guide for experimentalists interested in specifically labeling internal sequences on double-stranded (ds) DNA molecules for single-molecule experiments. We describe six labeling approaches demonstrated in a single-molecule context and discuss the merits and drawbacks of each approach with particular attention to the amount of specialized training and reagents required. By evaluating each approach according to criteria relevant to single-molecule experiments, including labeling yield and compatibility with cofactors such as Mg(2+), we provide a simple reference for selecting a labeling method for given experimental constraints. Intended for non-specialists seeking accessible solutions to DNA labeling challenges, the approaches outlined emphasize simplicity, robustness, suitability for use by non-biologists, and utility in diverse single-molecule experiments.
这篇综述是为有兴趣专门标记双链 (ds) DNA 分子内部序列以进行单分子实验的实验人员提供的实用指南。我们描述了在单分子环境中已经证明的六种标记方法，并讨论了每种方法的优缺点，特别关注所需的专门培训和试剂的数量。通过根据与单分子实验相关的标准评估每种方法，包括标记产率以及与镁 (Mg2+) 等辅助因子的兼容性，我们为根据给定的实验限制选择标记方法提供了一个简单的参考。本综述面向寻求 DNA 标记挑战的易于使用的解决方案的非专业人士，概述的方法强调了简单性、稳健性、适合非生物学家使用以及在各种单分子实验中的实用性。
